Isolation,Purification,Identification And Anti-Fatigue Activity Of Polysaccharides From Pleurotus Ostreatus

Pleurotus ostreatus was number two in the global commercial mushroom market,with the continuous optimization of cultivation conditions and market demand.Pleurotus ostreatus polysaccharide was one of the main active ingredients of Pleurotus ostreatus,it has numerous medicinal properties with functionalities in immune regulation,antitumor and antioxidant.Recently,its antifatigue activity has also attracted much attention from researchers.Multiple studies have demonstrated that body fatigue caused by oxidative stress,antioxidation was one of the main measures to relieve fatigue.In this paper,the structure of different components of Pleurotus ostreatus polysaccharides from two different areas will be characterized.The components with significant antioxidant activity was screened out by free radical scavenging ability.The antifatigue activity of Pleurotus ostreatus polysaccharide was verified by regulating antioxidant pathway in vivo and cellular tissue.The total polysaccharide?POPH and POPS?was extracted from the Pleurotus ostreatus by wall-broken method.By using DEAE-52 column chromatography,four purified polysaccharides?POPH-1,POPH-2,POPS-1 and POPS-2?were obtained.The structure of them was detected by using HPGPC,IC,FTIR and SEM.The free radical scavenging ability of them was compared.Constructed the L6 cells model of oxidative stress injury induced by H₂O₂,evaluated its protective effect on cellular tissue based on antioxidant by detecting cell activities,antioxidant enzyme activities,expression of MDA and ROS.The antifatigue activity of Pleurotus ostreatus polysaccharide was evaluated using loaded swimming test and biochemical indexes.The main results are as follow:?1?The yield of POPH and POPS was 4.3%and 6.6%.The molecular weight of POPH-1,POPH-2,POPS-1 and POPS-2 was 5.12×10⁵Da,3.98×10⁵Da,4.78×10⁵Da and 3.84×10⁵Da,they are all complex water-soluble polysaccharides mainly composed of glucose with high purity.POPH-2 and POPS-2 were pyranose with?glycosidically bound.The morphology of POPH-2 was irregular fragment with smooth surface and many holes.?2?POPH-2 and POPS-2 had significant antioxidant ability,the optimum of scavenging activity to DPPH and ABTS radicals were 71.6%and 92.2%respectively as the concentration of POPH-2 was 0.5-5mg/m L.POPH-2 protected L6 cells from injury induced by H₂O₂ by reducing the content of MDA?P<0.01?and ROS?P<0.01?,increasing the activities of SOD?P<0.05?and GSH-Px?P<0.05?.?3?POPH-2?200 mg/kg bw·d?can significantly extend swimming exhaustion time of mice.POPH-2 relieved fatigue of mice by reducing the content of BUN?P<0.05?,LDH?P<0.05?and LA?P<0.05?in serum,LA?P<0.05?and HG?P<0.05?in muscle,ROS in serum and liver,increasing the activities of SOD?P<0.01?in serum and GSH-Px?P<0.05?in liver.In summary,in this study,the structure of four components polysaccharide was characterized.POPH-2 was selected as an effective material through free radical scavenging experiment in vitro and antioxidant ability at cell level.The antifatigue activity of Pleurotus ostreatus polysaccharide was revealed preliminarily from its antioxidant level,which provided basis and strategy for the development of Pleurotus ostreatus polysaccharide and the research of antifatigue.